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CHPF2 / Chondroitin polymerizing factor 2, non-catalytic subunit · IHC design guide
Design Immunohistochemistry for CHPF2
Plan CHPF2 staining in paraffin sections using the reported cytoplasmic tissue pattern (HPA tissue IHC) and the catalog antibody’s 1:100–1:300 IHC dilution (datasheet). Compare positive and negative tissue compartments when interpreting staining (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CHPF2 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasmic staining (HPA tissue IHC); Golgi membrane protein (UniProt) | |
| Staining pattern | Cytoplasmic staining in most tissues (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 8.0 HIER, heat-mediated (datasheet A30609) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | Esophagus+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A30609) | |
| Caveat | Some squamous epithelia have no detectable staining (HPA tissue IHC) | |
| Regulation | Stimulus regulation unreported (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended CHPF2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A30609) is accompanied by a published colon cancer tissue protocol (PMC8377503 methods).
| Sample | Paraffin-embedded Human brain tissue; fixative not specified (datasheet A30609) |
| Fixation | Image fixative and duration unreported (datasheet A30609); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Tris-EDTA pH 8.0 (datasheet A30609); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-CHPF2, 1:100 - 1:300 (datasheet A30609) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CHPF2-positive staining in adipocytes of adipose tissue (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated Tris-EDTA pH 8.0 retrieval (datasheet A30609). The published protocol reports peroxide and serum pretreatment but gives no heat-retrieval buffer (PMC8377503 methods).
Section 2
What Is the Expected CHPF2 Staining Pattern?
In paraffin-section IHC, expect cytoplasmic CHPF2 staining in many cell types, including adipocytes, glandular cells, hematopoietic cells and glia (HPA: cytoplasmic expression in most tissues; listed cells Medium). A Golgi-associated component is plausible because CHPF2 is a membrane protein with a short cytoplasmic tail and a lumenal region (UniProt Q9P2E5: Golgi membrane; topology 1–6 and 30–772). The HPA tissue IHC profile is Approved, pending external verification (HPA: reliability description).
| Cytoplasmic chromogen in adipocytes, glandular cells, hematopoietic cells or glia. | This agrees with the reported compartment and sampled positive cells; the listed examples are Medium, so intensity need not be strong (HPA: tissue IHC). Assess identifiable cells and tissue architecture together. A compact perinuclear component can be compatible with Golgi residence, but HPA describes the tissue IHC pattern broadly as cytoplasmic (UniProt Q9P2E5: Golgi membrane; HPA: tissue profile). |
| Predominantly nuclear chromogen in a tissue section, with little cytoplasmic signal. | Treat this as discordant with the HPA tissue IHC profile and investigate section or detection artefact before assigning CHPF2 (HPA: cytoplasmic expression in most tissues). Nuclear signal is not a universal exclusion: HPA separately reports approved nucleoplasm and cytosol staining in ICC-IF, which uses a different specimen and readout (HPA: subcellular ICC-IF). |
| Strong staining in squamous epithelium of esophagus, oral mucosa or vagina. | These specific cell populations were Not detected in the supplied tissue IHC observations (HPA: esophagus, oral mucosa and vagina). Check whether the chromogen follows cells or a general tissue feature; possible explanations include antibody cross-reactivity or endogenous detection activity. The HPA observations are comparisons, not proof that every individual section must be blank. |
| Diffuse chromogen across cells, stroma and blank regions, obscuring cell boundaries. | A field-wide deposit cannot be scored reliably as the reported cytoplasmic pattern (HPA: tissue profile). In chromogenic IHC, nonspecific antibody binding, inadequate blocking or endogenous detection activity can raise background (standard IHC practice). Compare a reagent-control section and inspect whether any signal remains specifically associated with cells. |
| No visible signal in a sampled HPA Medium population. | An absent result in adipocytes or a listed glandular, hematopoietic or glial population conflicts with that particular HPA observation (HPA: tissue IHC, Medium), but does not alone show that CHPF2 is absent. Confirm that the relevant cells are present, then evaluate retrieval, antibody concentration, detection and controls using standard IHC practice; no CHPF2-specific retrieval condition is supplied. |
💡Expected CHPF2 appearanceCall a section supportive when identifiable HPA-positive cells show discernible cytoplasmic staining, often moderate in the listed examples; predominant nuclear staining, field-wide deposit or strong signal confined to HPA Not detected cell populations warrants review (HPA: tissue IHC; HPA: Approved, pending external verification).
| Subcellular context | Golgi membrane residence supports an intracellular expectation (UniProt Q9P2E5: subcellular location), while HPA tissue IHC reports cytoplasmic staining without resolving a Golgi-only pattern (HPA: tissue profile). Do not require a visible Golgi outline. |
| Topology and unknown epitope | CHPF2 has residues 1–6 cytoplasmic, 7–29 transmembrane and 30–772 lumenal (UniProt Q9P2E5: topology). The supplied record does not place the antibody epitope, so topology alone cannot specify retrieval or permeabilisation settings. |
| Isoforms and glycosylation | Two isoforms and glycosylation sites at residues 121 and 342 are annotated (UniProt Q9P2E5: isoforms; glycosylation). Without an epitope map or target-specific comparison, neither annotation predicts a different IHC staining pattern. |
| Evidence scope | The tissue antibody is IHC Approved, with external verification pending (HPA: reliability); its status is not Enhanced (HPA: antibody HPA020992, IHC Approved). No target-specific fixation sensitivity or antigen-retrieval effect is provided. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive cells appear blank in the paraffin section. | The run may have insufficient staining sensitivity, or the expected cells may be absent from the section; the supplied examples are Medium, not uniformly strong (HPA: tissue IHC). | Confirm cell identity and a working positive control. Review the catalog antibody's IHC-P instructions, retrieval, dilution and detection as general workflow checks; do not infer a CHPF2-specific fixation effect from HPA. |
| Chromogen spreads over the section or appears in reagent-control areas. | Background from nonspecific binding or endogenous detection activity can obscure a cellular result (standard chromogenic IHC practice). | Inspect the reagent control, blocking and detection chemistry; adjust one general workflow variable at a time, then rescore whether signal is confined to cells with cytoplasmic staining (HPA: tissue profile). |
| The strongest signal is nuclear in IHC. | That distribution differs from the reported tissue IHC pattern (HPA: cytoplasmic expression in most tissues). The approved nuclear ICC-IF observation cannot by itself validate nuclear chromogen in paraffin tissue (HPA: subcellular ICC-IF). | Compare with the cytoplasmic positive control and reagent control; document the discordance before interpreting the nuclear signal as CHPF2. |
| Squamous epithelial cells stain strongly while expected cells do not. | The supplied squamous populations are Not detected (HPA: esophagus, oral mucosa and vagina); cross-reactivity or endogenous detection activity is possible. | Recheck cell identity and controls, then compare another HPA Medium cell population in the same run (HPA: tissue IHC). Treat the unexpected pattern as unconfirmed. |
| Some tissues stain more weakly than the chosen positive control. | The tissue profile spans Medium, Low and Not detected observations, despite cytoplasmic expression in most tissues (HPA: tissue IHC). | Score the specified cell population against its reported level: for example, lung alveolar cells are Low, whereas adipocytes are Medium (HPA: tissue IHC). Avoid treating a weak Low population as a failed run. |
| Does an IF/ICC image need the same pattern as paraffin IHC? | The assays have distinct observations: HPA reports nucleoplasm and cytosol in ICC-IF, and a broadly cytoplasmic tissue IHC profile (HPA: subcellular ICC-IF; tissue IHC). | Interpret the IF/ICC result on its own guide page and keep this section's paraffin-section decision anchored to tissue IHC; the ICC status of HPA020992 is Approved (HPA: antibody validation). |
Experiment evidence
IHC & IF images showing CHPF2 staining
A30609 · IHCImmunohistochemical analysis of paraffin-embedded Human brain. Antibody was diluted at 1:100(4° overnight). High-pressure and temperature Tr
A14020-1 · IHCImmunohistochemistry validation of CHPF2 using Anti-CSGlcA-T CHPF2 Antibody (A14020-1). Immunohistochemical analysis of paraffin-embedded Hu
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000033100-CHPF2/tissue/cerebral+cortex#imid_5644496
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000033100-CHPF2/tissue/colon#imid_5644430
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000033100-CHPF2/tissue/liver#imid_5644431
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000033100-CHPF2/tissue/kidney#imid_5644416
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA020992; green = CHPF2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA020992; green = CHPF2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CHPF2 IHC & IF
🧪Run adipose tissue first and require staining in adipocytes (HPA: Medium in adipocytes), then run esophagus as a negative tissue and assess squamous epithelial cells (HPA: Not detected in esophageal squamous epithelial cells). On the adipose slide, inspect non-adipocyte areas for background, but do not assume those cells are CHPF2-negative without validation.
Positive control tissue: Adipose tissue (Adipocytes, HPA Medium)
Negative control tissue: Esophagus (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CHPF2 in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (approved), Cytosol (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; an isotype control matched to the primary antibody’s host species and clonality; and a biological specificity control such as CHPF2 knockout tissue or immunogen-peptide preabsorption (selected-SKU tissue-IHC caption: peptide-preabsorbed control). For chromogenic staining, quench endogenous peroxidase and check unstained adipose tissue for background before scoring.
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported; the selected-SKU paraffin-section caption does not state a fixative (selected-SKU tissue-IHC caption: fixative not stated). That caption reports high-pressure, high-temperature Tris-EDTA retrieval at pH 8.0 and antibody dilution 1:100 overnight at 4°C, but does not establish that retrieval is required (selected-SKU tissue-IHC caption). Whether frozen sections or IF are easier is unreported; in adipose sections, assess staining around lipid vacuoles cautiously because section morphology can make edge staining difficult to interpret.
HPA tissue IHC evidence for CHPF2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Pending external verification.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Vagina | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CHPF2 IHC Tips
For CHPF2 chromogenic IHC in paraffin sections, assess retrieval, compartment, and controls together before interpreting staining intensity.
How should I optimize retrieval when CHPF2 staining is weak in paraffin sections?
Start with heat-mediated Tris-EDTA at pH 8.0 for antigen retrieval (datasheet A30609). The paraffin-embedded human brain example used high-pressure, high-temperature retrieval with that buffer, followed by antibody at 1:100 overnight at 4 °C (caption A30609). If staining remains weak, vary heating time in small steps while keeping buffer, section thickness, antibody concentration, and detection conditions constant (standard IHC practice). Compare each condition with an adjacent section and a no-primary control, because stronger chromogen alone does not establish specific CHPF2 detection (standard IHC practice).
Could fixation explain inconsistent CHPF2 staining across paraffin blocks?
Target-specific fixation sensitivity is unknown: the selected paraffin-embedded brain caption does not state a fixative (caption A30609). Record the fixative and fixation duration for each block, then compare similarly processed sections before attributing intensity differences to CHPF2 expression (standard IHC practice). Use the stated Tris-EDTA retrieval at pH 8.0 and the reported 1:100, 4 °C overnight incubation as starting conditions for that antibody (datasheet A30609; caption A30609). If blocks differ in processing history, assess morphology and staining controls alongside CHPF2; neither tissue staining patterns nor CHPF2 topology establishes a fixation effect (standard IHC practice).
Which staining compartment is plausible for CHPF2, and how should discordant patterns be checked?
CHPF2 is annotated as a Golgi apparatus membrane protein with a cytoplasmic 1–6 region and a lumenal 30–772 region (UniProt Q9P2E5 topology and subcellular annotation). A perinuclear cytoplasmic pattern is therefore plausible in IHC, while HPA reports cytoplasmic expression in most tissues (UniProt Q9P2E5 subcellular annotation; HPA tissue IHC). HPA also reports approved nucleoplasm and cytosol locations from ICC/IF, so a nuclear signal needs assessment rather than automatic rejection (HPA subcellular). Check whether the pattern follows intact cell morphology, persists across adjacent sections, and exceeds no-primary background before assigning a compartment (standard IHC practice).
How can isoforms and membrane orientation affect CHPF2 epitope detection?
CHPF2 has 2 annotated isoforms, but the supplied antibody evidence does not locate its epitope or establish isoform coverage (UniProt Q9P2E5 isoforms; caption A30609). Its transmembrane segment spans residues 7–29, with residues 30–772 on the lumenal side, so epitope access may depend on where the antibody binds (UniProt Q9P2E5 topology). Glycosylation is annotated at residues 121 and 342; those sites alone do not establish antibody sensitivity to glycosylation (UniProt Q9P2E5 glycosylation). If staining differs between samples, confirm the antibody’s documented immunogen region before interpreting the difference as isoform-specific expression (standard IHC practice).
How should I follow up a CHPF2 IHC pattern with multiplex IF?
Use IF as a separate validation experiment, since the selected antibody evidence describes paraffin-section chromogenic IHC rather than an IF protocol (caption A30609). For cerebral cortex, pair CHPF2 with a marker appropriate to the glial subtype being assessed, because HPA reports medium staining in cortical glial cells (HPA tissue IHC). Choose fluorophores after checking unstained tissue autofluorescence and include single-label controls to assess spectral overlap (standard IF practice). Because the antibody epitope is unspecified, compare gentle permeabilisation conditions before interpreting an absent signal from the lumenal 30–772 region or the cytoplasmic 1–6 region (UniProt Q9P2E5 topology; standard IF practice).
What should I check when CHPF2 chromogenic staining is diffuse or patchy?
Run no-primary and peptide-competition controls where feasible; the selected brain image used antibody pre-absorption with immunogen peptide as its negative control (caption A30609; standard IHC practice). Check endogenous peroxidase blocking before DAB development and compare a shorter chromogen development with the original condition (standard chromogenic IHC practice). Examine section edges, folds, damaged areas, and precipitate before scoring diffuse color as cellular CHPF2 signal (standard IHC practice). Retain a documented positive section alongside each run; HPA reports medium CHPF2 staining in cerebral cortex glial cells, although its tissue IHC reliability is approved pending external verification (HPA tissue IHC).
How should I quantify CHPF2 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then use a consistent H-score from staining intensity and the percentage of positive cells (standard IHC practice). For spatial questions, report positive cells per mm² in prespecified regions, with the number of evaluable cells or tissue area as the denominator (standard IHC practice). Normalize comparisons to the same cell type, viable tissue area, staining run, and image acquisition settings (standard IHC practice). Report cytoplasmic and nuclear scores separately if both appear, because HPA tissue IHC describes predominantly cytoplasmic expression while HPA ICC/IF also reports nucleoplasm and cytosol locations (HPA tissue IHC; HPA subcellular).
When is a CHPF2-positive IHC result credible rather than artefactual?
Treat staining as more credible when it localizes to intact cells, exceeds no-primary background, and is reduced by immunogen-peptide competition under matched conditions (standard IHC practice; caption A30609). A perinuclear cytoplasmic pattern is compatible with Golgi membrane localization, while HPA tissue IHC describes cytoplasmic expression in most tissues (UniProt Q9P2E5 subcellular annotation; HPA tissue IHC). Interpret a predominantly nuclear pattern cautiously: HPA ICC/IF reports nucleoplasm localization, but that finding does not identify the source of a particular chromogenic signal (HPA subcellular; standard IHC practice). Exclude edge staining, necrotic regions, and residual endogenous enzyme activity before calling a cell positive (standard IHC practice).
Boster reagents
Best CHPF2 / Chondroitin polymerizing factor 2, non-catalytic subunit IHC Antibodies
Both catalog antibodies have paraffin-section IHC images from human brain (catalog image captions); IF is listed for both, and ICC for A30609 (catalog applications).
Real IHC data
Anti-CSGLCAT CHPF2 Antibody
Cat # A30609
Real IHC data
Anti-CSGlcA-T CHPF2 Antibody
Cat # A14020-1
A30609 has a human brain paraffin-section IHC image and lists Human, Mouse, and Rat reactivity plus IHC, IF, and ICC applications (A30609 image caption; catalog applications/reactivity). A14020-1 has a human brain paraffin-section IHC image and lists Human reactivity plus IHC and IF applications (A14020-1 image caption; catalog applications/reactivity).
Which to pick: For human brain paraffin-section IHC, either SKU has its own image at 1:100; A30609’s caption also reports Tris-EDTA retrieval at pH 8.0 and a peptide-absorbed control (A30609 and A14020-1 image captions). For IF/ICC, choose A30609 when ICC is needed because ICC and IF are listed for it, while A14020-1 lists IF only; neither has an IF image in the payload (catalog applications; IF image alts). For Mouse or Rat work, A30609 is the listed cross-species option and is described as polyclonal; both IHC captions show paraffin sections, with fixative unreported (catalog reactivity; A30609 dilution_raw; IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9P2E5 (CHPF2_HUMAN, Chondroitin polymerizing factor 2, non-catalytic subunit).
- Human Protein Atlas. CHPF2 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. CHPF2 subcellular location (ICC-IF): Localized to the nucleoplasm and cytosol..
- Human Protein Atlas. CHPF2 antibody validation summary (1 antibodies).
- Comprehensive Analysis of Glycolysis-Related Genes for Prognosis, Immune Features, and Candidate Drug Development in Colon Cancer. Frontiers in cell and developmental biology 2021 — PMC8377503.
- PubMed PMID:18316376 — UniProt-cited evidence.
- PubMed PMID:10718198 — UniProt-cited evidence.
- PubMed PMID:12975309 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CHPF2 (Chondroitin polymerizing factor 2, non-catalytic subunit). Expected localisation Cytoplasmic staining (HPA tissue IHC); Golgi membrane protein (UniProt). Antigen retrieval: Tris-EDTA pH 8.0 HIER, heat-mediated (datasheet A30609). Validated IHC antibody: A30609 at 1:100 - 1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative Esophagus. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded Human brain tissue; fixative not specified (datasheet A30609); 2. Fixation: Image fixative and duration unreported (datasheet A30609); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: Tris-EDTA pH 8.0 (datasheet A30609); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-CHPF2, 1:100 - 1:300 (datasheet A30609); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A30609); Caveat: Some squamous epithelia have no detectable staining (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unspecified (UniProt); Regulation: Stimulus regulation unreported (UniProt). Evidence: UniProt Q9P2E5, HPA tissue IHC · Approved, HPA ICC-IF · nucleoplasm and cytosol., Datasheet A30609, PMC8377503. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CHPF2/immunohistochemistry-ihc-guide.